Workshop Immuno-Imaging and Molecular Therapy

April 23th - 27th 2018

Auditorium TI. K Vrije Universiteit Brussel (VUB)Faculty of Medicine & PharmacyBuilding KLaarbeeklaan 1031090 BrusselsBelgiumBelgium

Contact:Email: carlien.geldof@gmail.com Phone: +32 2 477 49 24Website: http://www.icmibrussels.be/Courses.htmlFacebook: https://www.facebook.com/events/576311925908969/

WorkshopImmuno-ImagingandMolecularTherapy

Workshopdescription:

Duetotheirlowimmunogenicity,highaffinity/specificityandflexibilitytowardsbiotechnological engineering, antibodies are probably the most widely usedvehiclesformoleculardiagnosis,imagingandtherapy.Theworkshop‘Immuno-ImagingandMolecularTherapy’aimstobringtogethernationaland internationalexperts inthe fieldsofantibody-engineering,clinicaland preclinical nuclear imaging, tracer design, image acquisition andreconstruction,(radio)chemistry,targetedradionuclidetherapy,opticalimaging,immunogenicity and intellectual property. Altogether, a full overview of theresearch topic ‘immunotheranostics’willbeprovided through theorganizationof13lecturesand2half-dayhands-onsessions.Inparticular,aspecialfocuswillbe put on nanobodies as an example of engineered antibody vehicles for non-invasiveimagingandtargetedtherapy.Forwhom?

This workshop is targeted towards anybody interested in the broad topic of'immune-theranostics'. Basic knowledge of radiation, biotechnology, bio-chemistry,medicineandengineeringisrequired.MastersinBiology,BiomedicalSciences, Biotechnology, (Bio)Chemistry,Medicine or Pharmacy, PhD students,technologists and postdoctoral fellows working or interested in this field ofresearchareencouragedtoparticipate.Venue:

TheworkshopwillbeheldattheHealthCampusoftheFreeUniversityBrussels(VrijeUniversiteitBrussel),Laarbeeklaan103,B-1090Brussels,Belgium.Thelectureswillbeheldintheseminarroomofthedoctoralschool,buildingEground floor. Lunches, refreshments, discussions, practical sessions and thecentralmeetingpointwillbetheseminarroomoftheICMIlaboratory,buildingKgroundfloor(seehttp://www.icmibrussels.be/AboutUs/Directions.html)

Workshopschedule:

MondayApril23thtoFridayApril27th,2018,eachdayfrom9AMto5PM,unlessotherwisespecifiedAccommodation:

The workshop participants themselves are responsible for their ownaccommodation.Participantsseekingaccommodationcanbegivenadvice.Registrationdeadlineandfee:

April10th,2018€250for5-dayprogramRegistration includes attendance of the workshop, lunches, refreshments andparticipationtothesocialevent.Contact:

CarlienGeldofcarlien.geldof@gmail.comphone+32.2.477.4924OrganisationCommittee:

Director:ProfNickDevoogdtAdministration:CarlienGeldofProfMarleenKeyaertsProfSophieHernot

Program:

MondayApril239u00-10u00 ProfNickDevoogdt(VUB)

Welcome,roundtableintroduction10u00-11u30 ProfNickDevoogdt(VUB)

Do'sanddon’tsinimmunotracerdesign11u30-11u45 Bio-break11u45-12u45 ProfChristianVanHove(UGent)

Overviewofimagingmodalities,partI12u45-14u15 Sandwichlunch+postermounting14u15-15u15 ProfChristianVanHove(UGent)

Overviewofimagingmodalities,partII15u15-15u30 Bio-break15u30-17u00 ProfMarleenKeyaerts(VUB/UZBrussel) OverviewofclinicalnuclearmedicineTuesdayApril249u00-10u30 ProfSergeMuyldermans(VUB) Nanobodydiscoveryandbiotechnologicalapplications10u30-10u45 Bio-break10u45-12u15 ProfNickDevoogdt(VUB) Imagingwithnanobodiesinpreclinicalmodels12u15-13u45 Sandwichlunch+postersession113u45-15u15 ProfMarleenKeyaerts(VUB/UZBrussel) Clinicaltranslationofnanobody-tracers15u15-15u30 Bio-break15u30-17u00 ProfGeertRaes(VUB/VIB) Intellectualpropertyofbiologics17u00-... Socialevent(Brusselssightseeing&dinner)

WednesdayApril259u00-10u30 DrChloéAckaert(VUB) Immunogenicityofproteinsandnanobodies10u30-10u45 Bio-break10u45-12u15 ProfSophieHernot(VUB) Fluorescenceandcardiovascularapplicationsofnanobodies12u15-13u30 sandwichlunch+postersession213u30-17u00 Hands-onsession(tobechosenfromoneofthetopics)

-Analysis of nanobody in vivo biodistribution data in mice(Dr.MatthiasD’Huyvetter)-nanobodyleadselectionanalysis(ProfNickDevoogdt)-visittothenuclearmedicinedepartmentandcyclotronunitoftheUZBrusselhospital:dailypatientcareandclinicaltrialprocedures(ProfsMarleenKeyaerts&Caveliers)-Opticalimaginginmice(ProfSophieHernot)-Nanobodyradiochemistrytechniques(ProfCatarinaXavier)

ThursdayApril269u00-10u30 DrGéraldineGebhart&DrZénaWimana(Bordet Institute Brussels) Clinicalimmuno-PET10u30-10u45 Bio-break10u45-12u15 ProfMatthiasD'Huyvetter(VUB) Targetedradionuclidetherapywithnanobodies12u15-13u30 sandwichlunch+postersession313u30-17u00 Hands-on session (to be chosen from one of the topics

indicatedabove)

FridayApril279u00-10u30 DrAnnieVanBroekhoven(Q-Biologicals) ProteinCMC10u30-10u45 Bio-break10u45-12u15 ProfBrianZeglis(HunterCollege,NewYork,USA) InVivoPretargeting:RadiosynthesisattheTumorSurface12u15-13u00 Sandwichlunch13u00-14u30 ProfJean-PierrePouget(ICRM,Montpellier,France) Targetedandoff-targeteffectsofradionuclidetherapy14u30-15u30 Round-tablediscussion,workshopconclusion&group

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Informationaboutthelecturers:

NickDevoogdt

ProfessorVrijeUniversiteitBrussel(VUB)ResearchclusterImagingandPhysicalSciences(BEFY)InvivoCellularandMolecularImaginglaboratoryndevoogd@vub.ac.bePhone:+3224774991

Nick Devoogdt was trained as a Master in Molecular Biology at the Free University ofBrussels (Vrije Universiteit Brussel, VUB), where he obtained his degree with highdistinctionin1997.HeobtainedanFWOscholarshipandin2004obtainedaPhDinAppliedBiological Sciences under the promotorship of Prof De Baetselier, working on cancergenetics, cellular biology and immunity. Again he acquired a FWO grant to continue hispostdoctoralresearchfortheperiod2004-2007.In2004-2005hewasavisitingscientistatthe National Cancer Institute in Bethesda USA, studying ovarian cancer genetics andbiomarkers, after which he returned to VUB in Belgium for additional training as apostdoctoralscientist:in2005-2006inthefieldofcellularimmunologyandcancergenetics,in2007inantibody-engineeringandfrom2008onwardsinmolecularimaging.In2013hebecameassistantprofessorinthesmallanimalimaginglabICMIandin2016fullprofessor.In 2014 he also co-founded the spinoff company Camel-IDS where he is still active ascoordinatorofthediscoverypipeline.

His research aims to develop novel applications in molecular and nuclear imaging andtargeted therapies.His current focus is on the nanobody-technology as targeting vehicles.More in particular, his research is focused on the generation of new probes for theirapplication (nuclear or other types of imaging and therapy) in small animal models ofdisease,andinasecondphaseontestinginpatients.Hence,hispreclinicalandtranslationalresearchissupportiveandcloselyconnectedwithclinically-orientedresearchteamswithinthefacultyofmedicineandtheuniversityhospital.

ChristianVanhove

AssociateprofessorGhentUniversityMedicalImagingandSignalProcessingHeadofpreclinicalcoreimagingfacilityUGent(INFINITY) Christian.Vanhove@UGent.bePhone:+3293324115

ChristianVanhovegraduatedasaBiomedicalandClinicalEngineerfromBrusselsUniversityin1990.From1991until1996,heworkedasaMedicalPhysicist at theNuclearMedicinedepartment of the Sint-Elisabeth hospital in Zottegem, where he was doing research anddevelopments forthe industry inaclinicalenvironment.Researchanddevelopmentswerefocusedonall aspectsofmedical imageprocessing, including image reconstruction, imageregistration and image quantification. In 1996, he moved to the Nuclear MedicinedepartmentoftheBrusselsUniversityHospital.AsaMedicalPhysicistExperthecontinuedhis research in the field of medical image processing and obtained his PhD in MedicalScience in 2004. During his PhD, a new algorithm for the automatic segmentation andquantification of gated blood pool SPECT images, in both humans and small animals,wasdevelopedandvalidated. In2005,hewasoneof the initiatorsof thesmallanimal imaginglab of Brussels University and worked as a postdoctoral researcher at this preclinicalimagingfacility.DuringthisperiodhisresearchwasfocusedonquantitativepinholeSPECT.Since February 2011, he joined the Medical Imaging and Signal Processing (MEDISIP)researchgroupoftheFacultyofEngineeringofGhentUniversity,whereheisresponsibleforthe INnovative Flemish IN-vivo Imaging TechnologY (INFINITY) lab. This lab focuses onmultimodal in-vivo imaging strategies and serves as a core facility for preclinical imagingwithin Ghent University. The major research domains of INFINITY are the evaluation ofpathophysiology in neurological diseases, cancer and inflammation. Christian Vanhove isassociateprofessorandlecturerinmolecularimaging.

MarleenKeyaerts

AssistantprofessorVrije Universiteit Brussel InVivoCellularandMolecularImagingLaboratory (ICMI)HeadofClinicsUZBrussel–DepartmentofNuclearMedicine Marleen.Keyaerts@vub.ac.be Phone:+3224775020

MarleenstudiedmedicineattheVrijeUniversiteitBrusselandperformedhermasterthesisat the nuclear medicine department of the UZ Brussel on the biodistribution of a newradiopharmaceutical in healthy subjects. She immediately startedher PhD at the ICMI labafter graduation,whichwas completed in 2011. In her PhD project,Marleen investigatedbioluminescenceimagingfortheassessmentoftumorburden.Whenshemovedbacktotheclinictocompleteherclinicaltraininginnuclearmedicine,afirstnanobodyleadcompoundwasreadyfor first inhumantesting.Marleenwasstrongly involved inthisphaseIclinicaltrial using anti-HER2 Nanobody PET/CT imaging in breast carcinoma patients and she iscurrentlyprimaryinvestigatorinthephaseIItrial.

SergeMuyldermans

ProfessorVrijeUniversiteitBrusselLaboratoryofCellularandMolecularImmunologysvmuylde@vub.ac.be Phone:+3226291969

Serge Muyldermans started his studies (chemistry) at the ‘Vrije Universiteit Brussel’ andobtainedhisPhDin1982onstructuralaspectsofnucleosomesandchromatin.Asapostdocat the same university he became involved in the early 90-ies in the camel antibodyprogram.In2003hewasappointedProfessorattheVrijeUniversiteitBrussel,whereheisnow heading the camel antibody engineering group in the laboratory of Cellular andMolecularImmunology.ThediscoveryofcamelHeavy-chainantibodiesandtheapplicationsoftheiruniquesinglevariableantigen-bindingdomain(nowreferredtoasNanobodies)ledto the publication of over 170 articles in peer reviewed international journals, many ofwhichappeared in top journals. In January2002,hewasco-founderof ‘Ablynx’, a spin-offcompany that generates Nanobodies for therapeutic purposes. Ablynx is currentlyemploying~300peopleandhasseveralNanobodiesinclinicaltrials.

GeertRaes

Assistantprofessor&valorisationmanager VUBCellularandMolecularImmunologyunitStaffScientistatVIBInflammationResearchCentre Geert.Raes@vib-vub.be Phone: +3226291968

Afterobtainingadegreeinbio-engineeringandaPhDintumorimmunology,Geerthasbeenstaff scientist at VIB, where he has been involved in projects aimed at translating basicimmunologyresearchintoindustrialapplications.Since2013,hehasapartialappointmentasvalorisationmanageratVrijeUniversiteitBrussel, focusingoneconomicvalorisationofnanobodyapplications inmolecular imagingand therapy.Geert is co-inventoron6patentfamilies. He is co-founder and Head of IP and Contracting of the VUB spin-off companyCamel-IDS. ChloéAckaert

PostdoctoralresearcheratVrijeUniversiteitBrussel–Cellularandmolecularimmunologylaboratorychloe.ackaert@vub.ac.be Phone: +3226291978

Chloé studied Pharmacy at the Katholieke Universiteit Leuven and performed hermasterthesisattheHôpitalBichatinParis,theCHUinCharleroiandtheUZAntwerpenonEnzymeReplacementTherapy inFabrydiseasepatients.Shedidan internship in the laboratoryofTherapeuticandDiagnosticAntibodies(KUL)ontheImmunogenicityofanti-TNFamAbsinIBD patients, followed by a PhD on the immunogenicity of modified allergens at theUniversityofSalzburginAustria.AfterherPhD,shewasinvolvedinthestart-upoftheCROImmunXperts,offeringintegratedimmunogenicityservices.Since2015,ChloéisworkingintheCamelAntibodygroupwithintheCellularandMolecularImmunologylab(VUB)whereshefocussesonstudyingtheimmunogenicityofNanobodies. Sheperformedtheanti-drugantibodymeasurements inthephaseIclinical trialusinganti-HER2NanobodyforPET/CTimaginginbreastcarcinomapatientsandsheiscurrentlyassessingtheimmunogenicityriskofseveralNanobodiesbyinvitromethods.

SophieHernot

AssistantProfessorVrijeUniversiteitBrusselInvivoCellularandMolecularImagingLaboratorySophie.hernot@vub.ac.bePhone: +3224774991

Sophie Hernot is a Bio-engineer from the VUB. She completed her PhD thesis inmedicalsciences in 2011 on the use of microbubbles (ultrasound contrast agents) as well asnanobodies for molecular imaging and drug delivery applications. As post-doctoralresearcher, and currently as assistant professor in the laboratory of In vivo Cellular andMolecularImaging(ICMI,VUB),herresearchinterestsaretwo-fold:1)designandvalidationof nanobodies for optical imaging applications, and in particular towards a clinicaltranslationforimage-guidedsurgery,and2)validationofnanobody-basedmolecularprobesforidentificationandcharacterizationofhigh-riskatheroscleroticlesions.

MatthiasD’Huyvetter

PostdoctoralresearcherattheVrijeUniversiteitBrusselInVivoCellularandMolecularImagingLaboratorymdhuyvet@vub.ac.be Phone: +3224774991

MatthiasstudiedBiomedicalSciencesattheUniversityofAntwerp,andobtainedhisMasterdegree in 2009. From 2009 to 2013 he received a SCK•CEN/VUB PhD scholarship that,together with a one-year research grant of the foundation Emmanuel van der Schueren(2013-2014),ledtoaPhDthesisentitled:“RadiolabeledNanobodiesasTheranosticToolsinCancer Treatment”. One year of his PhD was conducted at the Duke University MedicalCentreinDurham,USA,withthesupportoftheBelgianEducationalFoundation(BAEF).InOctober2014,heco-foundedtheVUBspin-offcompanyCamel-IDSNV/SA.Currently,heisaprofessorattheVUBandholdsapostdoctoralmandateofFWOFlanders.

CatarinaXavier

AssistantprofessoratVrijeUniversiteit BrusselInVivoCellularandMolecularImagingLaboratory(ICMI)cxavier@vub.ac.be Phone:+3224774991

CatarinastudiedChemistryattheUniversityofLisbon,andobtainedherdegreein2002.Sheimmediately started working at ITN/Portugal and in 2004 she was awarded with a PhDgrant from the Portuguese Science Foundation (FCT). Her PhD studies (2004-2009)wereperformedat ITN/UniversityofLisbonundersupervisionofDr. IsabelRegodosSantos,atUniversityofZurichunder thesupervisionofDr.RogerAlbertoandatUniversityofMilanundersupervisionofDr.CleliaGiannini.AsfromApril2009sheisapostdoctoralresearcherat the In Vivo Cellular and Molecular Imaging Laboratory. In 2015 she became assistantprofessoratICMI.Her research aims at the development of new radiopharmaceutical probes for imaging ortargeted radionuclide therapy. She is involved in the process of nanobody probeoptimizationandtranslationtotheclinicforPET/CTimaging.

GeraldineGebhartMD,PhD,PhysicianinNuclearMedicine,JulesBordetInstitute,Brussels geraldine.gebhart@bordet.be

GeraldinestudiedmedicineattheUniversitéLibredeBruxelles,includingoneyearinBerlinaspartoftheErasmusprogram.In 2009, with a good background of internal medicine, she started to work in NuclearMedicine under the supervision of Prof. Patrick Flamen. She discovered with greatenthusiasm, molecular imaging, which is today an emerging field in oncology. Her PhDprojectfocusedonthecontributionofmolecularimagingtoearlyevaluationofresponsetoanti-HER2agentsinBreastCancer.Inbrief,shecontributedtodifferentprojectsincluding- The evaluation of the FDG-PET/CT scans performed in the biological window of theNEOALTTO trial, which has compared, in HER2 positive breast cancer patients, thepreoperativeadministrationoftrastuzumab, lapatinibortheircombination(paclitaxelwasaddedsubsequentlytoeachofthese3arms)-Studieswithzirconium-89labelledtrastuzumabinpatientswithHER2positivemetastaticbreastcancer:theZEPHIRstudy,aphaseIIprospectiveimagingstudyevaluatingtheutilityof pre-treatment zirconium-89 labelled trastuzumab PET/CT and an early FDG-PET/CTresponse to identifypatientswithadvancedbreastcancerunlikely tobenefit fromanovelanti-HER2:T-DM1.TheworkofG.Gebhartwasrecognizedbythefollowingawards:JNMEditor'sChoiceAward2013andAlaviMandellAwardforJNMarticlepublishedin2013.

ZénaWimana

PhD,BiomedicaldoctorandcoordinatoroftheradiopharmacyNuclearMedicine/Radiopharmacy,JulesBordetInstitute,Brusselszena.wimana@bordet.beZéna studied Biomedical Sciences at the University of Antwerp. She then concluded hertrainingwithapostgraduatedegreeinBiomedicalImagingpartiallyperformedattheVUB,whereshestayedoneyearasascientificassistant.In2009,shestartedtoworkasaresearchassociateintheNuclearMedicinedepartmentofProf. Patrick Flamen at the Jules Bordet institute. She started a research project one yearlaterunder the supervisionofProf.GhanemGhanemandobtainedherPhD from theULBwithathesisentitled“ImmunoPetimagingusingZirconium-89radiolabeledtrastuzumabtoexploreresistanceinHER2+/MUC4+breastcancer”.In the mean time she became the coordinator of the radiopharmacy of the Jules BordetInstituteandparticipatedtotheintroductionofseveralradiopharmaceuticalsintheclinicaspremier in Belgium, namely 89Zr-Trastuzumab for the imaging of HER2 positive breastcancer,177Lu-DOTATATEforthetherapyofneuro-endocrinetumorsand68Ga-PSMAfortheimagingofprostatecancer.She is currently also a boardmember in different scientific and professional associations,includingtheBelgianNuclearMedicineSociety,BelgianMolecularImagingCommunityandtheRadiopharmacyboardoftheNationalInstituteofRadioelements.

AnnieVanBroekhovenCEOatAmatsiQBiologicalsa.vanbroekhoven@amatsigroup.com Annie Van Broekhoven PhD, graduated as a Master in Chemistry from the University ofAntwerp(Belgium)andobtainedaPhDin1981.AfterapostdocattheWeizmannInstitute,(Rehovot,Israel)inthegroupofProf.Dr.U.Littaueronthecloningoftubulinandactin,shebecame involved in the expression, fermentation, purification and manufacturing ofindustrialenzymesforPetrofina(Total).ShejoinedInnogeneticsin1992,firstasresponsibleforthebioprocessdepartment,laterasVice-PresidentBiologicalsresponsible for theprocessandanalyticalmethoddevelopment,and production of Biologicals for diagnostic and therapeutic usage, including cGMPmanufacturing.ShehasbeenamemberoftheboardofdirectorsofRheinBiotechNVandoftheVRWB(FlemishBoardofScientificPolicy).Currently she is a member of the steering committee of LABIP (Lactic Acid BacterialIndustrialPlatform)andRIOFI(IndustrialCouncilofUniversityAntwerp).OnaregularbasissheservesasanexternalexpertfortheFlemishInstituteofScienceandTechnology(IWT)and European Framework Programs. She is a Professor at The University of Antwerp,teachingIndustrialBiotechnologyandMicrobiology.

BrianZeglis

Assistant Professor of Chemistry, Hunter College of the CityUniversityofNewYorkAttending Radiochemist (Affiliate), Department of Radiology atMemorialSloanKetteringCancerCentreAssistant Professor (Adjunct), Department of Radiology at WeillCornellMedicalCollege.

bz102@hunter.cuny.edu Phone:212-896-0443

Dr. Zeglis received his training at Yale University (advisor: Robert H. Crabtree), the CaliforniaInstitute of Technology (advisor: Jacqueline K. Barton), and Memorial Sloan Kettering CancerCentre (advisor: Jason S. Lewis). The Zeglis Laboratory of Hunter College is composed of 3postdoctoralfellows,5graduatestudents,and5undergraduatestudentsandisdedicatedtothesynthesis, validation, and clinical translation of radiopharmaceuticals for the imaging andtherapyofcancer.

Jean-PierrePouget

Headofthe“RadiobiologyandTargetedRadiotherapy”teamofIRCMINSERM,Montpellier,France

jean-pierre.pouget@inserm.frPhone:+33467613708

Dr Jean-Pierre Pouget obtained his PhD thesis in Radiobiology in 2000 from the CurieInstitute in Paris and carried-out a post-doctoral fellowship in the Nuclear MedicineResearch Laboratory at Barts and the London School ofMedicine andDentistry. He thenmovedtoParis totheFrenchRadiationProtectionandNuclearSafetyAgency(IRSN)for4yearswhereheworkedonradiationcasualties. AftermovingtoMontpellier,hejoinedtheFrenchNationalInstituteforHealthandMedicalresearch(INSERM)whereheisnowleaderof the Radiobiology and Targeted Radiotherapy team at the Cancer Research Institute ofMontpellier (INSERM, France). He develops new radiopharmaceuticals for cancer imagingandtherapywithaspecialfocusonradiobiology.Hehaspublishedabout60papersdealingwith radiobiologyand radionuclide therapyand severalpatents.Besides researchactivity,DrPougetisinvolvedinteachingattheUniversityofMontpellier.

Informationabouttheprogram:

Theartofmakinganimmunotracer(NickDevoogdt)In thispresentation, the concept ‘molecular imaging’will be introducedanda generaloverviewofdifferenttypesoftracerswillbegiven.Amajorfocuswillbeonantibody-tracers: howdoes the immune systemdevelops an efficient antibody response?Whatare the molecular and structural features of antibodies? What are the differenttechniquestogenerateantibodiesandhowcantheybeengineeredintodifferenttypesof antibody-fragments? The pharmacokinetic behaviour of antibodies and engineeredfragmentswill be explained, and their relation tomolecular andnuclear imaging.Theantibodypretargetingapproachwillbebriefly touchedupon.Finally,variousconceptsrelated to tracer design will be explained, including ‘affinity’, ‘avidity’, ‘enhancedpermeability and retention’, ‘the site-barrier-effect in tissue penetration’, ‘specificactivity’and‘antigenicsink’.Thisfirstlecturewillalsotrytoprovidelinksandanintroductiontootherlecturesgivenduringtheworkshop.Overviewofimagingmodalities(ChristianVanhove)In 1895 the German physicist Wilhelm Conrad Roentgen discovered the X-rays, anachievement that earned him the first Nobel Prize in Physics in 1901. These X-raysproducedthefirstmedicalimagesinthebeginningofthepreviouscentury.Nowadays,awiderangeofimagingtechniquesareavailablethatcanberoughlysub-dividedintotwomain categories: structural and functional imaging devices. Themost commonly usedstructural imaging technologies are computed tomography (CT), magnetic resonanceimaging(MRI)andultrasoundimaging(US).Thesetechniquesweredevelopedtonon-invasively visualize nonspecific macroscopic anatomical and physiological changes intissues. Functional imaging modalities, such as single photon emission tomography(SPECT), positron emission tomography (PET) and optical imaging (OI) focus on thevisualizationofmolecular/cellulartargetsinlivingsubjects.Thesetargetsinfunctionalimagingcanincludetransporters,cellsurfacereceptorsandintracellularenzymes.An overview will be presented of the physical basics behind the image formationprocess of these imaging modalities. The requirements to move from clinical topreclinicalimagingwillalsobediscussedforeachmodality.ThepresentationwillstartwithCT that canbe seenas adirect evolutionofX-ray imaging. Secondly, thenuclearimagingtechniquesSPECTandPETwillbepresented,includingtheimportantevolutionof combining these nuclear imaging techniques with CT andMRI. Themore compleximageformationprocessofMRIwillbeshortlyintroduced.Finally,OIwillbepresented,including bioluminescence and fluorescence in-vivo imaging, which are of moreimportanceinthepreclinicalarena.

Overviewofclinicalnuclearmedicine(MarleenKeyaerts)Thispresentationwillgoovertheroutinelyperformednuclearmedicineexaminationsin patients, with a focus on the uptake mechanism of different tracers as well as apersonal view on the future of clinical nuclear medicine. The session is intended forstudents and researchers in the preclinical field that would like to get a betterunderstanding of daily practice in a nuclear medicine ward in the hospital. Typicalimagesofroutinelyperformedscanswillbediscussedandinterpreted.Nanobodydiscoveryandbiotechnologicalapplications(SergeMuyldermans)Llama and camels have unique antibodies comprising a homodimer of heavy chainsonly, whereby the antigen is recognized by virtue of one single domain. Astraightforward technology was developed to immunize a camelid, to clone therepertoireofantigen-bindingfragments,fromwhichtheantigen-specificfragmentsareidentified after phage display selections. The resulting recombinant, antigen-bindingsingle-domainantibodyfragmentsarealsoreferredtoasNanobodies(Nbs)becauseoftheirsizeof4nmby2.5nmindiameter.Nanobodies are well produced in microbial systems, very robust and highly soluble,bind their cognate antigenwith high affinity and specificity. Very often theNanobodyrecognizes an epitope that is difficult to targetwith human ormouse antibodies. The‘humanization’ofacamelderivedsingledomainantibodyisstraightforward.OneofthelargestadvantageofNanobodiescomesfromtheirstrictmonomericbehavior,theeasetotailorthemintolargerpluripotentconstructsandtheirfunctionalitywhenexpressedintracellularly.Such beneficial properties of Nanobodies over other antigen-binding fragments fromconventionalantibodiesinspiredmanyresearcherstoemployNanobodiesasaversatiletoolinvariousinnovativeapplicationsinbiotechnologyandmedicineas:

• aresearchtooltoimmune-capturetheantigenfromcomplexmixtures,• apotentprobetotrace(oreliminate)targetantigenwithinlivingcells,• anexcellentdiagnostictoolfornon-invasiveinvivoimagingoftumors,• atherapeutictooltoeradicatetumorsortrypanosomeinfectioninmicemodels,• ananti-venomtoprotectfromscorpionenvenoming

Imagingwithnanobodiesinpreclinicalmodels(NickDevoogdt)Inthislecture,thefocuswillbeonnanobodiesandtheirevaluationbynuclearimagingin animal models of various diseases. The lecture will start with a rather detailedoverview of protocols to generate good nanobody vehicles and will then look atsubsequentattentionpointsonwhichparametersandcriteriashouldbeconsideredtoselectaleadcompound.Wewilllookatstructuralfeaturesofnanobodiesandhowthisisreflectedintheiraminoacidsequence.Themajorpartofthislecturewillconsistofanoverview of recent published and unpublished applications of nanobodies in smallanimal nuclear imaging in pathologies ranging fromoncology to autoimmunedisease,macrophage-tracking,andsomeexamplesofnanobodyengineeringtoamelioratetracerperformance.Clinicaltranslationofnanobody-tracers(MarleenKeyaerts)Theclinical translationofnewimagingagentswillbediscussedusingthe68-Ga-HER2nanobodyasanexample.Participantswillget insight intoregulatoryrequirementsforclinical trials, including GMP compliance, clinical trial applicationwith the competentauthorities and ICH-GCP requirements. A study protocol for a phase I study will bepresentedandexplained,aswellastheobtainedresults.Intellectualpropertyofbiologics(GeertRaes)Using nanobodies as an example, some basic principles of intellectual property rightswillbediscussed.Afterabriefoverviewof themain featuresand typesof intellectualproperty rights, the focuswill be on patents: advantages of patents, requirements forpatentability,limitationstopatentablesubjectmatterandformulationofpatentclaims.Finally,thedistinctionbetweenpatentabilityandfreedom-to-operatewillbeexplained.Immunogenicityofrecombinantproteins(ChloéAckaert)The immunogenicity of recombinant biopharmaceuticals will be discussed with thefocus on clinical impact of immunogenicity and methods available to assess theimmunogenicity risk. Participants will get insight into in silico, in vitro and in vivomethodsforimmunogenicityriskassessment,aswellasanti-drugantibodymonitoringin clinical trial phases. Case studies will be discussed as illustration and theimmunogenicityoftheGa-NOTA-anti-HER2Nanobodywillbepresented.

Fluorescenceandcardiovascularapplicationsofnanobodies(SophieHernot)In vivo Fluorescence Imaging - A general overview will be given regarding theapplicationoffluorescenceimagingforimage-guidedsurgery.Boththetechnicalaspectsofopticalimagingdevicesaswellasthedesignoffluorescenttracerswillbediscussed.Current and emerging clinical applications will be illustrated, as well as the role thenanobodiesinthisfield.Cardiovascularmolecular imaging -This talkwill focuson the rolemolecular imagingcanplay in thediagnosisandcharacterizationof cardiovasculardiseases, inparticularforvulnerableatheroscleroticplaques.Besidesanoverviewof thecurrentstateof theart,theimportantpitfallsandhurdlesthatthefieldhastofacewillbecovered.Finally,the contributionofnanobodies for imagingof atherosclerotic lesionsand follow-upofinflammationaftermyocardialinfarctionwillbecovered.ClinicalimmunoPET(Z.Wimana&G.Gebhart)Currently, mAb-based imaging forms the main category of immunoPET, largely as aresultofthedevelopmentanduseofmAbsasdisease-selectivesystemictreatmentsofcancer.ImmunoPETcombinesthesensitivityandhighspatialresolutionofPETwiththespecificityofhighaffinityligandsofimmuneorigin.It canbeused fordifferentapplication includingcharacterizationof lesions,diagnosis,staging,evaluationandpredictionoftreatmentresponse.Furthermore,inR&DofmAb-based treatment, immunoPET can offer valuable information on in vivo drug-targetinteraction, pharmacokinetics and pharmacodynamics, when the drug itself isradiolabeled and used as an imaging probe. As for the use in the characterisation oftumors, immunoPET presents several advantages: it is a real time whole-body non-invasive tool, allowing simultaneous cancer detection and localization by visuallyproving the presence of an expressed biomarker, both in the primary tumor and inmetastases.Moreover,itprovidesanattractivenovelalternativeinvivooptionastoIHCstaining, improving diagnostic and tumor characterization aswell as heterogeneity oftarget expression. In thisway, immunoPET could influence treatment choice and thiscouldresultinimprovedpatient’soutcomes.Targetedradionuclidetherapy:Nanobodiesasanexample(MatthiasD’Huyvetter)ThispresentationwillintroducethebasicsofTargetedRadionuclideTherapyofcancer,andbrieflyhighlightdifferentexcitingapplicationsthatarenowavailable intheclinic.Inaddition,animportantpartofthetalkwillfocusontheuseofNanobodiesaspotentialvehicles for Targeted Radionuclide Therapy using both beta-- and alpha-particleemittingisotopes.

InVivoPretargeting:RadiosynthesisattheTumorSurface(BrianZeglis)Invivopretargetingstandsasapromisingapproachtoharnessingtheexquisitetumor-targetingpropertiesofantibodiesfornuclearimagingandtherapywhilesimultaneouslyskirting their pharmacokinetic limitations. The core premise of pretargeting lies inadministering the targeting vector and radionuclide separately and having the twocomponentscombinewithinthebody.Inthismanner,pretargetingstrategiesdecreasethecirculationtimeoftheradioactivity,reducetheuptakeoftheradionuclideinhealthynon-target tissues, and facilitate the use of short-lived radionuclides that wouldotherwise be incompatible with antibody-based vectors. In this lecture, we seek toprovideabriefsurveyof the fourpreeminentmechanisticapproaches topretargeting,strategies predicated on streptavidin and biotin, bispecific antibodies, complementaryoligonucleotides, andbiorthogonal click chemistry.Particular attentionwill bepaid topretargetingbasedontherapidandbiorthogonalinverseelectron-demandDiels-Alderreactionbetweentetrazineandtrans-cyclooctene.TargetedandOff-TargetEffectsOfRadionuclideTherapy(Jean-PierrePouget)For long, radiation biology has relied on the “target theory“ that considers that cellshavetobehitbyionizingparticlestobekilled.Incancertherapy,theprobabilityofhitspercellasafunctionoftheradiationdosewastheninitiallydeterminedbythePoissonlaw and next was assessed using linear quadratic mathematical models. Radiationharmfuleffectsaremostlydescribedbyinteractionofradiationwithwater,whichisthemajorconstituentoflivebeings,andnextwithnuclearDNA,whichcontainsthegeneticinformationhasbeenthecenterofmostofResearch.Theyinvolveclusteredproductionofradicalsandreactiveoxygenspecies(ROS)(O2-°.,HO°,H2O2).However, in the last twenty years, many studies have highlighted the role ofextranuclear subcompartments such as mitochondria or cell membrane. Moreover,signals originating from irradiated cells have also been shown to be responsible forcytotoxic and genotoxic effects in non-irradiated neighboring cells. The so-calledbystander effects would include sustained production of ROS and reactive nitrogenspecies (RNS) in neighboring cells but also of cytokines, ATP and extracellular DNAmaking the bystander response similar to inflammation. Also, immune cells could berecruitedandactivatedbydangerassociatedmolecularpatternsreleasedbyirradiatedcellsandparticipatetodistanteffectsofirradiation(called“abscopal”effects).Because of the physical characteristics of radionuclide therapy (low dose rate andprotracted exposure, low and high LET particles, heterogeneous and whole bodyirradiation) these new data have to be considered in nuclear medicine for bettertreatmentefficacyandbetterriskassessment.

InformationabouttheHands-Onsessions

AnalysisofNanobodyinvivobiodistributiondatainmice(MatthiasD’Huyvetter) During this practical session, participants will be able to get acquainted with thedifferentstepsinvolvedwhenevaluatingthebiodistributionofaradiolabeledNanobodyinananimalmodel.ThesessionstartswiththeradiolabelingofaNanobody,followedbythe intravenous injection of the sample into amouse, after which amicro-SPECT/CTimagewillbegenerated.Finally,wewillanalyzethegeneratedmicro-SPECT/CTimagesusingAMIDEsoftware.Nanobodyleadselectionanalysis(NickDevoogdt)In this practical session, I will give an overview of various techniques andmethodologies that are important in the lead selection analysis of nanobodies. Thismeansthatyouwillbeabletobetterunderstandwhichparameterscanbelookedattofind the best nanobody for your diagnostic or therapeutic purpose. Usually, whenperforming biopanning of a phage-displayed immune library, several candidatenanobodiescanbeidentified,whichcanbesubjectedtoanumberofteststorankthemon best performance. Techniques that will be looked at are ELISA, surface-plasmonresonance, DNA and amino acid sequence analysis, Thermofluor assays and flowcytometry,andproductionandpurificationofnanobodies fromE.colicultures.Duetolackoftime,theseexperimentswillnotbeperformedassuch.Instead,experimentswillbesimulatedandexamplesofthematerialandequipmentusedwillbeprovided.Visitandoperationsofhospitalnuclearmedicinedepartment(MarleenKeyaerts)During this practical session, participants will visit the different parts of the nuclearmedicinedepartmentintheUZBrussel,includingradiopharmacyandcyclotronfacility,patient areas and cameras, clinical workstation areas. Afterwards, the procedures,paperwork and practical requirements to perform clinical trials with newradiopharmaceuticals will be explained using the Nanobody-based clinical trials asexamples.Finally,participantswillhave thechance topersonallyreviewand interpretPET/CTimagesofpatientsthatparticipatedinthediagnosticnanobodytrials.Image-guided surgery using near-infrared fluorescently-labeled nanobodies(SophieHernot)This session will begin with a guided tour of the fluorescence imaging platform,equipped with several optical systems for in vivo macroscopic imaging (2D/3D) incombinationwithinvitroandexvivohighresolutionimagingdevices.The purpose of the practical course given in this session is to gain insight into theimportance of a well-designed imaging probe with an appropriate biodistributionprofile,aswellastoexperiencethebenefitofimage-guidedsurgeryinadiseasemodel.Nanobodyradiochemistrytechniques(CatarinaXavier)Duringthispracticalsession,participantswillbeabletolearnhowtolabelNanobodieswith 99mTcand68Ga.All thestepsof theradiolabeling,qualitycontrol,purificationandfinalformulationwillbeaddressed.

Registration Form - Immuno-Imaging and Molecular Therapy

Brussels (Belgium) - April 23-27 2018 First Name: _________________________________________________________________ Family Name: _________________________________________________________________ Date of Birth (dd/mm/yyyy): _________________________________________________________________ Place of Birth: _________________________________________________________________ Nationality: _________________________________________________________________ Gender: o Female o Male Telephone: (+___)____________________________________________________________ GSM: _________________________________________________________________ E-mail: _________________________________________________________________ --------------------------------------------------------------------------------------------------------------------------------------- This section is not obligatory for Master Students following the official VUB Course ‘Molecular Imaging’: Institution / Company Name: _________________________________________________________________ Laboratory / Research Group: _________________________________________________________________ Address: Street:______________________________________________ Nr:__________ Postal Code:______________ City:____________________________________ Country:__________________________________________________________ Position: o Master Student o PhD Student o Postdoctoral Fellow o Other: Specify __________________________________________________ Research Focus: _________________________________________________________________ Participants are expected to bring along a poster describing their research topic(s), which will be discussed during the walking through poster sessions -------------------------------------------------------------------------------------------------------------------------------------- I will register as o an external participant and pay the registration fee of € 250 o a Master/PhD student of the VUB Masterclass ‘Molecular Imaging’ (Official registration) This registration fee includes the workshop attendance, lunches, refreshments and social event. The registration fee is not applicable for Master/PhD Students following the official VUB Course ‘Molecular Imaging’. I will attend the social event: o Yes o No For students following the official VUB course ‘Molecular Imaging’ we would like to receive a payment of € 25 for the organisation of the social event. I prefer the following hands-on sessions: 1. ____________________________________________________________________________________ 2. ____________________________________________________________________________________ 3. Back-up: ____________________________________________________________________________ Hands-on Sessions: - Analysis of nanobody in vivo biodistribution data in mice (Dr. Matthias D’Huyvetter) - Nanobody lead selection analysis (Prof. Dr. Nick Devoogdt) - Visit and operations of hospital nuclear medicine department (Prof. Dr. Marleen Keyaerts) - Image-guided surgery using near-infrared fluorescently-labeled nanobodies (Prof. Dr. Sophie Hernot) - Nanobody radiochemistry techniques (Prof. Dr. Catarina Xavier) All registrations must be submitted by email to carlien.geldof@gmail.com. The invoice will be provided after receiving the completed registration form. Your registration will be valid after receiving a proof-of-payment of the registration fee.